IMMUNOLOGICAL MONITORING The core was initiated at the beginning of the last funding period. It provides tissue processing services and immune monitoring assays for cancer center members. The daily operations are under the supervision of Joel Whitfield who is the manager of the core. Two full-time research associates perform the core functions. During the last four years, 24 different users have utilized the service. Over 19,500 samples have been processed/assayed during this period. Recharges are developed for individual users to cover costs of laboratory supplies and labor. The core services are listed below. Services: Tissue processing and cryopreservation. This unique service involves isolation of peripheral blood monocytes (PBMC) or plasma from blood samples (i.e., blood draws or aphaeresis products); cryopreservation of PBMC or plasma/serum; tumor dispersion of solid tumors into single cell suspensions; cryopreservation of tumor samples; RNA or DNA extraction of tissue samples. These services to not duplicate the services of the Tissue Core. Storage of cryopreserved tissue samples Staining of PBMCs with monoclonal antibodies for phenotypic analysis by FACS (analysis to be provided by Flow Cytometry Core located in close proximity to the Immunology Core) ELISA assays of cytokines (murine and human IFN-y, GM-CSF, IL-10, IL-4, IL-6, TNF-a) Lymphocyte proliferation assays (i.e., KLH, tumor cells, mutagens) Mixed lymphocyte reaction (MLR) assays ELISPOT assays of cytokines (murine and human): The core has an automated spot reading for the ELISPOT assay. It also has the software necessary to analyze ELISPOT data which allows investigators to quantitate how many lymphoid cells secrete a specific cytokine in a response to antigen on a per cell basis. Lymphocyte cytokine release assays to antigens (i.e., tumor antigen or KLH) Lymphocyte cytoxicity assays to tumor targets (i.e., 4-hour chromium release assay) Quantitative RT-PCR for cytokine mRNA analysis of tissue samples.